Printing needle holder device, sterile enclosure and method of printing and culturing biological cells

Abstract

A sterile enclosure including a container and a printing needle holder device fixed to the container. The container having a base wall, a side wall, the side wall and the base wall delimiting an interior chamber, one inlet port and outlet port and an aperture. The printing needle holder device having a needle adapter, a printing needle and a flexible sheath having a first end edge fixed to the needle adapter and a second end edge fixed to the aperture of the container, a three dimensional biological structure being intended to be printed on the base wall, the interior chamber being intended to receive a cell culture media.

1 - 23 . (canceled) 
     
     
         24 . Printing needle holder device adapted to be fixed to a container having an aperture, said printing needle holder device comprising:
 at least one needle adapter,   at least a printing needle extending in a defined direction, the at least one needle adapter holding the printing needle,   a flexible sheath having a first end edge tightly fixed to the at least one needle adapter and a second end edge delimiting an opening intended to be fixed to the aperture of the container,   the printing needle holder authorizing the displacement of the printing needle in at least two perpendicular directions when the second end edge is fixed to the container for printing a three-dimensional biological structure in the container, the two perpendicular directions being perpendicular to the defined direction.   
     
     
         25 . Printing needle holder device adapted to be fixed to a container having an aperture, said printing needle holder device comprising:
 at least one rigid frame and a septum supported by the rigid frame, the septum being designed to be punctured by a printing needle,   a flexible sheath having a first end edge tightly fixed to the at least one rigid frame and a second end edge delimiting an opening intended to be fixed to the aperture of the container,   the printing needle holder authorizing the displacement of the printing needle in at least two directions when the second end edge is fixed to the container for printing a three-dimensional biological structure in the container, the two perpendicular directions being in a plane parallel to the septum.   
     
     
         26 . Printing needle holder device according to  claim 24 , which comprises a rigid flange fixed to the second edge, the rigid flange being intended to be fixed to the aperture of the container. 
     
     
         27 . Printing needle holder device according to  claim 26 , wherein the rigid frame is provided with a first connector part intended to be fixed to the container. 
     
     
         28 . Printing needle holder device according to  claim 24 , wherein the at least one flexible sheath is made of one material among a thermoplastic elastomer, a thermoset and a silicone. 
     
     
         29 . Printing needle holder device according to  claim 24 , which comprises a receiving sleeve adapted to receive the needle. 
     
     
         30 . Sterile enclosure comprising a container said container comprising:
 a base wall,   at least one side wall, the at least one side wall and the base wall delimiting an interior chamber,   at least one inlet port and at least one outlet port,   an aperture,   characterized in that it comprises a printing needle holder device comprising:   at least one needle adapter,   at least a printing needle extending in a defined direction, the at least one needle adapter holding the printing needle,   a flexible sheath having a first end edge tightly fixed to the at least one needle adapter and a second end edge delimiting an opening,   a rigid flange fixed to the second edge; the printing needle holder authorizing the displacement of the printing needle in at least two perpendicular directions when the second end edge is fixed to the container for printing a three-dimensional biological structure in the container, the two perpendicular directions being perpendicular to the defined direction;   wherein the rigid flange of the printing needle holder device is fixed to the container to close the aperture; the three dimensional biological structure being intended to be printed on the base wall, the interior chamber being intended to receive a cell culture media.   
     
     
         31 . Sterile enclosure comprising a container, said container comprising:
 a base wall,   at least one side wall, the at least one side wall and the base wall delimiting an interior chamber,   at least one inlet port and at least one outlet port,   an upper rigid flange delimiting an aperture,   characterized in that it comprises a printing needle holder device ( 2 , 36 , 42 , 58 ) fixed to the aperture of the container, said printing needle holder device comprising:   at least one needle adapter,   at least a printing needle extending in a defined direction, the at least one needle adapter holding the printing needle,   a flexible sheath having a first end edge tightly fixed to the at least one needle adapter and a second end edge delimiting an opening fixed to the aperture of the container,   the printing needle holder authorizing the displacement of the printing needle in at least two perpendicular directions when the second end edge is fixed to the container for printing a three-dimensional biological structure in the container, the two perpendicular directions being perpendicular to the defined direction the upper rigid flange being fixed to an opening of the printing needle holder device, the three dimensional biological structure being intended to be printed on the base wall, the interior chamber being intended to receive a cell culture media.   
     
     
         32 . Sterile enclosure according to  claim 30 , wherein the base wall and the at least one side wall are made with a flexible film. 
     
     
         33 . Sterile enclosure according to  claim 30 , wherein the base wall is rigid and the at least one side wall is made with a flexible film. 
     
     
         34 . Sterile enclosure according to  claim 30 , wherein the base wall is made with a flexible film and the at least one side wall is rigid. 
     
     
         35 . Sterile enclosure according to  claim 30 , wherein the base wall and the at least one side wall are rigid. 
     
     
         36 . Sterile enclosure according to  claim 30 , which comprises a receiving sleeve for receiving the printing needle. 
     
     
         37 . Sterile enclosure according to  claim 30 , which comprises a vessel for priming the flow of bioink in the printing needle and/or for purging the printing needle. 
     
     
         38 . Sterile enclosure according to  claim 30 , wherein the base wall comprises at least one extension protruding from the base wall in a horizontal plane. 
     
     
         39 . Sterile enclosure according to  claim 38 , wherein the extension comprises reliefs to reference the location of the enclosure with respect to a support. 
     
     
         40 . Sterile enclosure according to  claim 30 , wherein the rigid flange of the printing needle holder device is fixed to the upper rigid flange of the container. 
     
     
         41 . Sterile enclosure according to  claim 40 , which comprises a septum fixed to the upper rigid flange. 
     
     
         42 . Method of printing and culturing a three dimensional biological structure in a sterile enclosure conform to  claim 30 , the method comprising the successive following steps:
 printing of the three dimensional biological structure, the flexible sheath authorizing the move of the needle adapter and the printing needle in at least two perpendicular directions during the biological printing step while keeping the sterility of the interior chamber,   supplying a cell culture media to the interior chamber.   
     
     
         43 . Method of printing according to  claim 42  which further comprises a fixation of the flexible sheath, for example by welding, to separate the printing needle from the interior chamber. 
     
     
         44 . Printing needle holder device according to  claim 25 , which comprises a rigid flange fixed to the second edge, the rigid flange being intended to be fixed to the aperture of the container. 
     
     
         45 . Printing needle holder device according to  claim 44 , wherein the rigid frame is provided with a first connector part intended to be fixed to the container. 
     
     
         46 . Printing needle holder device according to  claim 25 , wherein the at least one flexible sheath is made of one material among a thermoplastic elastomer, a thermoset and a silicone. 
     
     
         47 . Printing needle holder device according to  claim 25 , which comprises a receiving sleeve adapted to receive the needle. 
     
     
         48 . Sterile enclosure according to  claim 31 , wherein the base wall and the at least one side wall are made with a flexible film. 
     
     
         49 . Sterile enclosure according to  claim 31 , wherein the base wall is rigid and the at least one side wall is made with a flexible film. 
     
     
         50 . Sterile enclosure according to  claim 31 , wherein the base wall is made with a flexible film and the at least one side wall is rigid. 
     
     
         51 . Sterile enclosure according to  claim 31 , wherein the base wall and the at least one side wall are rigid. 
     
     
         52 . Sterile enclosure according to  claim 31 , which comprises a vessel for priming the flow of bioink in the printing needle and/or for purging the printing needle. 
     
     
         53 . Sterile enclosure according to  claim 31 , wherein the base wall comprises at least one extension protruding from the base wall in a horizontal plane. 
     
     
         54 . Sterile enclosure according to  claim 53 , wherein the extension comprises reliefs to reference the location of the enclosure with respect to a support. 
     
     
         55 . Sterile enclosure comprising a container; said container comprising:
 a base wall,   at least one side wall, the at least one side wall and the base wall delimiting an interior chamber,   at least one inlet port and at least one outlet port,   an aperture,   characterized in that it comprises a printing needle holder device comprising:   at least one rigid frame and a septum supported by the rigid frame, the septum being designed to be punctured by a printing needle,   a flexible sheath having a first end edge tightly fixed to the at least one rigid frame and a second end edge delimiting an opening,   a rigid flange fixed to the second edge, the printing needle holder authorizing the displacement of the printing needle in at least two directions when the second end edge is fixed to the container for printing a three-dimensional biological structure in the container, the two perpendicular directions being in a plane parallel to the septum; and wherein the rigid flange of the printing needle holder device is fixed to the container to close the aperture; the three dimensional biological structure being intended to be printed on the base wall, the interior chamber being intended to receive a cell culture media.   
     
     
         56 . Sterile enclosure according to  claim 55 , wherein the base wall and the at least one side wall are made with a flexible film. 
     
     
         57 . Sterile enclosure according to  claim 55 , wherein the base wall is rigid and the at least one side wall is made with a flexible film. 
     
     
         58 . Sterile enclosure according to  claim 55 , wherein the base wall is made with a flexible film and the at least one side wall is rigid. 
     
     
         59 . Sterile enclosure according to  claim 55 , wherein the base wall and the at least one side wall are rigid. 
     
     
         60 . Sterile enclosure according to  claim 55 , which comprises a receiving sleeve for receiving the printing needle. 
     
     
         61 . Sterile enclosure according to  claim 55 , which comprises a vessel for priming the flow of bioink in the printing needle and/or for purging the printing needle. 
     
     
         62 . Sterile enclosure according to  claim 55 , wherein the base wall comprises at least one extension protruding from the base wall in a horizontal plane. 
     
     
         63 . Sterile enclosure according to  claim 62 , wherein the extension comprises reliefs to reference the location of the enclosure with respect to a support.